The process using a synthetic library that generates multiple diverse human single domain antibodies

Area of Science:

• Biotechnology
• Immunology
• Drug Discovery

Background:

• Single domain antibodies (sdAbs) offer unique advantages for developing advanced therapeutics.
• Their specific binding characteristics are crucial for targeting disease-related antigens.

Purpose of the Study:

• To develop a method for generating single domain antibodies (sdAbs) against disease-relevant antigen conformations.
• To demonstrate the utility of these sdAbs in creating differentiated bispecific and multispecific antibody formats.

Main Methods:

• Utilized a fully synthetic phage display library based on a human IGHV3 scaffold for antibody generation.
• Employed diverse phage biopanning techniques against target antigens, followed by next-generation sequencing (NGS) and automated cloning.
• Assessed antibody binding via various assays, including flow cytometry, cell internalization, and activation assays.

Main Results:

• Successfully generated sdAbs (VHOs) capable of binding to disease-relevant antigen conformations.
• Identified multiple VHO sequences with distinct epitope specificities and activities through comprehensive screening.
• Demonstrated the expression of VHOs as Fc fusion proteins and their potential in bispecific and multispecific antibody formats.

Conclusions:

• A fully synthetic phage display library is effective for generating sdAbs targeting specific antigen conformations.
• Screening assays can identify distinct sdAb activities, enabling the development of novel therapeutic antibody formats.